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Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (11): 1242-1247.doi: 10.3736/jcim20111113

• Original Experimental Research • Previous Articles     Next Articles

Crohn disease in rats induced by different concentrations of trinitrobenzenesulfonic acid and ethanol

Shu Yang, Xin-yue Wang(), Shan Jing, Xue Yang, Yi-hua Sheng, Xin Yan   

  1. Department of Digestive Diseases, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
  • Received:2011-06-28 Accepted:2011-08-01 Online:2011-11-20 Published:2011-11-15

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Objective: To induce Crohn disease in rats by intraluminal instillations of different concentrations of 2,4,6-trinitrobenzenesulfonic acid (TNBS) and ethanol.
Methods: Crohn disease in rats was induced with enema containing TNBS and 50% ethanol with volume ratio of 2∶1 (experimental group 1) or 1∶1 (experimental group 2), or solution containing TNBS and anhydrous ethanol with volume ratio of 2∶1 (experimental group 3) or 1∶1 (experimental group 4). Equivalent volume of normal saline was used to set as the normal saline control, and rats without any treatment were set as the normal control group. The rats were killed at various time points (3, 7, 14 and 21 d) respectively. Colonic inflammation and damage were assessed macroscopically and histologically.
Results: In the colon of rats in the experimental group 3, discontinuous erosion, ulceration and infiltration of neutrophils occurred after one week; pebble sign and even segmental inflammation appeared on day 14. On day 21, it appeared improvement in the colon tissue and obviously thickened bowel wall, but the inflammation was easily observed under the light microscope. Experimental group 1 was similar to experimental group 3 in appearance of the colon on days 3 and 7; on day 14, colonic inflammation and damage were improved as compared with the experimental group 3, but there were obvious individual differences in histological findings among rats in group 1. In the experimental group 2, the intestinal wall turned to be normal on day 14. In the experimental group 4, there were high mortality and extensive damage of colon tissues, and the pathological characteristics were quite different from Crohn disease in humans.
Conclusion: The characteristics of the rat model of Crohn disease induced with a enema containing TNBS and anhydrous ethanol with volume ratio of 2∶1 are similar to the clinical features of human Crohn disease, including typical pathological characteristics and long duration of inflammation. It may be an ideal experimental model for studying pathogenesis of Crohn disease and for evaluating treatment effects.

Key words: Crohn's disease, disease models, animal, trinitrobenzenesulfonic acid, ethanol, rats

Table 1

Body weight of rats in each group at different time points (x±s, g)"

Day 1 Day 3 Day 7 Day 14 Day 21
Group n Body weight n Body weight n Body weight n Body weight n Body weight

Normal control		 30
171.767±5.367
6
182.333±2.251
6		 212.500±5.244
9
262.444±6.616
9		 336.111±5.862

Saline control		 30
172.600±5.360
6
185.000±4.472
6		 213.333±7.528
9
260.556±8.457
9		 334.778±6.648

Experimental 1		 30
171.333±5.371
6
161.333±
3.830**△△
6		 140.833±
2.041**△△▲▲■■
8
183.125±
3.720**△△▲▲□□■■
9		 245.333±
5.895**△△▲▲□□■■

Experimental 2		 30
171.467±5.964
6
162.167±
4.708**△△
6		 191.667±
4.082**△△□□■■
9
233.333±
4.330**△△□□■■
9		 303.667±
5.788**△△□□■■

Experimental 3		 30
171.400±5.411
6
161.667±
4.502**△△
6		 141.667±
2.582**△△■■
7
173.571±
3.505**△△■■
8		 211.250±
2.315**△△■■

Experimental 4		 30
170.800±5.307
6
160.500±
5.244**△△
6		 133.333±
4.082**△△
6
153.667±
6.218**△△
6		 187.667±
4.546**△△

Table 2

Microscopic evaluation of rats in each group at different time points (x±s)"



Group
Day 3
Day 7		 Day 14 Day21
n
Macroscopic grade
n
Macroscopic grade
n		 Macroscopic grade n Macroscopic grade

Saline control
6
0.183±0.117
6
0.100±0.110
9
0.056±0.073
9		 0.033±0.050

Experimental 1
6
8.333±
1.080△△▲▲■■
6
7.500±
1.095△△▲▲■■
8
6.250±
0.655△△▲▲□□■■
9
5.167±
0.500△△▲□□■■

Experimental 2
6
7.000±
0.775△△□□■■
6
5.750±
0.987△△□□■■
9
5.111±
0.220△△□□■■
9
4.722±
0.441△△□□■■

Experimental 3
6
8.667±
0.516△△
6
7.767±
0.408△△■
7
7.657±
0.690△△■
8
6.575±
0.403△△■

Experimental 4
6
9.417±
0.376△△
6
8.750±
0.274△△
6
8.250±
0.274△△
6
7.083±
0.376△△

Table 3

Histological evaluation of rats in each experimental group at different time points (x±s)"


Group		 Day 3 Day 7 Day 14 Day 21

n
Histological grade
n
Histological grade
n
Histological grade
n
Histological grade

Saline control
6
0.083±0.098
6
0.050±0.055
9
0.056±0.053
9		 0.056±0.073

Experimental 1
6
5.667±
0.683△△▲■■
6
5.250±
0.274△△▲■■
8
4.638±
0.619△△□□■■
9		 4.222±
0.441△△□■■

Experimental 2
6
5.000±
0.447△△□□■■
6
4.667±
0.516△△□□■■
9
4.456±
0.442△△□□■■
9		 3.800±
0.557△△□□■■

Experimental 3
6
6.133±
0.432△△■■
6
5.650±
0.288△△■■
7
5.457±
0.408△△■■
8		 4.750±
0.598△△■■

Experimental 4
6
7.500±
0.775△△
6
7.417±
0.585△△
6
6.767±
0.606△△
6		 5.500±
0.447△△

Figure 1

Changes of intestinal tissues in each group observed by hematoxylin and eosin staining on day 14 after modeling (Light microscopy, ×200) A: Experimental group 1; B: Experimental group 2; C: Experimental group 3; D: Experimental group 4; E: Saline control group."

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